1.Expression of Notch1, Jagged1 and NICD in epithelial ovarian carcinomas and a preliminary study on the activity of gamma-secretase in epithelial ovarian carcinoma cell lines
Zhaoyi FENG ; Hongwu WEN ; Qinping LIAO
Chinese Journal of Obstetrics and Gynecology 2014;49(10):780-786
Objective To study the expression of Notch 1,Jagged1 and Notch intracellular domain (NICD) in epithelial ovarian carcinoma tissues and analyze the clinical significance.To explore the activity of γ-secretase in epithelial ovarian carcinoma cell line SKOV3 and the effect of N-[N-(3,5-dil uorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester (DAPT),a γ-secretase inhibitor on the activity of γ-secretase in SKOV3.Methods Immunohistochemistry staining method was performed in 43 patients with epithelial ovarian carcinoma and 11 patients with benign epithelial ovarian tumor to detect the expression of Notch1,Jagged1 and NICD.The differences of expressionof Notch1,Jagged1 and NICD between malignant and benign ovarian tumors was compared and alsoanalyzed the correlation with clinicopathological parameters of ovarian carcinoma.Human serous ovarian cancer cell line SKOV3 and immortalized nontumorigenic ovarian epithelial cell line T29 were incubated in vitro.The activities of γ-secretase in SKOV3 and T29 with dimethyl sulfoxide (DMSO) and DAPT were detected respectively by Gal4VP16/UAS and dual luciferase reporter assay system.Results (1) The immunohistochemical composite scores (ICS) of Notch1 in epithelial ovarian carcinoma (6.7±2.2) were not significantly different with those in benign epithelial ovarian tumor (5.4± 2.7,P=0.153),while the ICS of Jagged 1 and NICD in epithelial ovarian carcinoma (5.3± 2.4,5.3± 2.3) were higher than those in benign epithelial ovarian tumor (1.6± 1.4,3.1± 1.7; all P<0.01).The expression of Notch 1,Jagged 1 and N ICD had no correlation with patients' aged,history of carcinoma,ascites,the level of serum CA125,maximum length of ovarian tumor,Federation International of Gynecology and Obstetrics (FIGO) stage,grade and pathology subtypes (all P>0.05).The hazard ratio between the high expression of Notch1,Jagged1,or NICD and the moderate to low expression of Notch1,Jagged1,or NICD,and Jagged1 were 0.771,1.648 and 1.316,respectively (all P>0.05).The 5-year survival rate and median survival time between the high expression of Notch,Jagged 1 or NICD in subgroup and moderate to low expression in subgroup were of no difference (all P>0.05).The activity of γ-secretase in SKOV3 was significantly higher than that in T29 [(12.2± 1.4)%,P=0.019].(2)After DAPT treated,the relative activity of γ-secretase in SKOV3 (50 μmol/L) was declined from (100.0±5.3)% to (6.6±0.8)% (P=0.001).Conclusions Jagged1 and NICD in Notch1 pathway may play a key role in the occurrence of ovarian carcinoma.The activity of γ-secretase in epithelial ovarian carcinoma was higher than that in ovarian epithelial cell which suggest that DAPT,γ-secretase inhibitor,may become the target of ovarian carcinoma treatment.
2.Pharmacokinetic study of iguratimod in rats
Feng XIAO ; Zhaoyi YANG ; Shangxue YAN ; Wei WEI
Chinese Journal of Primary Medicine and Pharmacy 2008;15(6):-
ObjectiveTo study the pharmacokinetics of iguratimod in rats. MethodsThe concentration ofiguratirnod in the samples was determined by HPLC method. The pharmacokineties parameters were calculated withDAS softwrare. ResultsThe mainpharmacokineties parameters of normal group(6mg/kg) were as follows:t1/2Ke:3.56h, tpeak: 4.00h, Cmax : 8.87μg/ml, AUC0.24 : 74.76μg· ml-1·h-1. The main pharmacokineties parameters of threemodel groups(3,6,12mg/kg) were as follows: t1/2Ke: 4.54,3.20,3.17h, tpcak:3.83,3.83,4.67h,Cmax:3.84, 8.31,12.69μg/ml, AUC0.24 :40.21,76. 72,117.06μg·ml-1·h-1. Except Cmax and AUC, no significant differenceswere found between the three model groups. And the differences between normal group and model group were notsignificant. ConclusionThe pharmacokinetics of rats ks fit to one-compartment model.
3.Study on shell shape changes of filial generation Oncomelania hupensis snails in Weishan Lake region,Shandong Province
Feng MIAO ; Xin LIU ; Lilei WANG ; Xuli DENG ; Xixin CHEN ; Zhaoyi FU ; Yongbin WANG
Chinese Journal of Schistosomiasis Control 2014;(1):13-15
Objective To explore the shape change characteristics of Oncomelania hupensis snail shell after the snails being passively migrated into Shandong intake area of the South-to-North Water Diversion Project. Methods The snails raised on the Dushan island in Weishan Lake region were captured,and 115 the first filial generation snails and 107 the second filial generation ones were selected. The length and width of shells and apertures,and the labial ridge thickness of those snails were measured,the longitudinal rib number of snail spiral was counted,and 107 parental snails were chosen as controls. Results The labial ridge thickness of the filial generation snail was significantly reduced with the increase of algebras,and the labial ridge thickness among the 3 generations had a significant difference(P<0.01). Both the length of snail shell and the width of snail apertures were de-creased,while the width of snail shell,the length of snail apertures,the multiplication product of snail aperture’s length and width and the longitudinal rib number of snail spiral were all increased. Conclusion The body size of Oncomelania snails in Weishan Lake region becomes smaller,and their shells become thinner,which indicates that the environment of the lake region is not suitable for snail breeding,and the snails have natural decay tendency with the extension of time.
4.Construction of lentiviral vector-mediated siRNA knockdown of ER-α36 and its action on gastric cancer cell growth
Xuming WANG ; Xuan HUANG ; Zhengqi FU ; Feng ZOU ; Shangkun ZHANG ; Zhaoyi WANG ; Lijiang LIU
Chinese Journal of Pathophysiology 2014;(12):2113-2119
[ ABSTRACT] AIM:To construct a lentiviral vector for stable delivery of the ER-α36 gene and to detect its effect on SGC7901 cell growth.METHODS: The efficient RNAi targeting sequences identified for the ER-α36 gene were screened.The Oligo DNA was synthesized with target sequences and annealed to form double-stranded DNA.Then it was digested by Xho I and EcoR I and connected with GV307 vector to produce LV-ER-α36-RNAi lentiviral vector.PCR was used to screen the positive clones and sequence.The LV-ER-α36-RNAi, pHelper 1.0 and pHelper 2.0 plasmids were co-transfected into 293T cells for producing lentiviral vector and infecting SGC7901 cell line.Fluorescence microscopy, real-time PCR and Western blotting were used to detect the transfection efficiency and gene silencing effect.17β-estrodial at concentration of 1 ×10 -10 mol/L was used to stimulate the recombinant cell line, and the action on the growth of gastric cancer cells and the expression of Src, ERK1/2 and cyclin D1 were determined.RESULTS: DNA sequencing analysis confirmed the identity of recombinant shRNA expression vectors.Immunofluorescence assay demonstrated that transfection efficiency was above 80%.Transfection of LV-ER-α36-RNAi significantly knocked down the expression of ER-α36 at mR-NA and protein levels with tetracycline ( TeT) simulating as revealed by real-time PCR and Western blotting.Compared with control group, the growth of the recombinant cell line declined and the expression of Src, ERK1/2 and cyclin D1 and the activation of Src decreased (P<0.05).CONCLUSION: Lentiviral vectors that silence ER-α36 expression are con-structed successfully and can be used to study the role of ER-α36 in gastric cancer.The ER-α36 is related with many kinds of cancer cell growth, including gastric cancer cells.
5.Clinical observation of green laser vaporization for the treatment of bladder tumor in 522 patients
Zhihua ZHANG ; Yatong CHEN ; Zhaoyi LI ; Tingji ZHANG ; Qiqing FENG ; Jian LI
Tianjin Medical Journal 2017;45(9):976-979
Objective To investigate the safety and efficacy of photoselective green laser vaporization of bladder tumor (PVBT). Methods A total of 522 patients with bladder tumor were enrolled in present study from January 2010 to May 2015, including 405 cases of non muscle-invasive bladder cancer (NMIBC) and 117 cases of muscle-invasive bladder cancer (MIBC). All of patients were treated with PVBT and intravesical instillation of epirubicin. Patients with MIBC received intravenous chemotherapy (kisi-hama and cisplatin). Results The hospitalization time was (7.32±1.28) days, the operation time was (27.08±5.36) min, and the indwelling urinary catheter was (2.42±0.34) days for patients in NMIBC group. During the follow-up period (12-60 months), 38 cases (9.4%) relapsed, of which 3 cases underwent radical cystectomy, and other 35 cases underwent PVBT again. All 405 patients were alive at the end of follow-ups. The hospitalization time was(26.18 ± 1.92) days, the operation time was (38.32 ± 6.54) min, and the time of indwelling urinary catheter was (2.72 ± 0.85) days for patients of MIBC group. During the follow-up period (12-60 months), 19 cases (16.2%) relapsed. Among them, 4 patients underwent radical cystectomy, and other 15 cases underwent PVBT. Six patients died from distant organ metastasis (including 2 cases of pulmonary metastasis and 4 cases of bone metastasis), and other 111 patients survived. Conclusion PVBT is safe and effective in the clinical application, especially for NMIBC and MIBC patients who are unable or unwilling to undergo radical cystectomy.
6.Investigation on schistosome infection and KAP about schistosomiasis control among boatmen in Weishan Lake region, Shandong Province
Yongbin WANG ; Feng MIAO ; Qingkuan WEI ; Zhaoyi FU ; Changlei ZHAO ; Xixin CHEN ; Xin LIU ; Yunhong YIN ; Changjun MA ; Zhongxue ZHANG ; Zhaocai FANG ; Juan WANG ; Changyin WEI
Chinese Journal of Schistosomiasis Control 2009;21(6):544-546
To understand the possibility of schistosomiasis transmission on the East Route Project of the South-to-North Water Diversion Project, a survey of endemic status of schistosomiasis in Weishan Lake area was conducted. A cluster sampling of 2086 boatmen were screened with DDIA Kit, and the positive ones were examined with Kato-Katz technique. Meanwhile, a questionnaire survey about schistosomiasis control was carried out to collect the data about the boatmen's general information and knowledge , attitude and practice (KAP). The results showed there were no schistosomiasis patients. However, there exist potential risk factors for schistosomiasis transmission because of the bad hygiene habits and the poor knowledge of schistosomiasis prevention in boatmen.